The HUVEC line was purchased from PromoCell (Heidelberg, Germany) (#C-12200). Frozen HUVECs were thawed and cultured according to the manufacturer’s protocol. Upon confluency, the HUVECs were detached by 15 min incubation with trypsin–EDTA (0.25%) (Invitrogen, Waltham, MA, USA) and seeded onto culture dishes at a density of 500,000 cells/cm2 in Endothelial Cell Growth Medium 2 (Ready-to-use) (PromoCell, #C-22011) supplemented with Endothelial Cell Growth Medium 2 SupplementMix (PromoCell, #C-39216) (hereafter referred as “culture medium”). Cells from passages five to eight were used in the recellularization experiments. Cell morphology was microscopically checked, and cultures with abnormal cell shapes were excluded from further experiments.
Endothelial cell growth medium 2 supplementmix
Manufactured by PromoCell
Sourced in United States
Endothelial Cell Growth Medium 2 SupplementMix is a supplement designed to support the growth and maintenance of endothelial cells in culture. It contains essential components required for endothelial cell proliferation and function.
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Lab products found in correlation
Huvecs, by Thermo Fisher Scientific (1 mentions)
Trypsin edta 0.25, by Thermo Fisher Scientific (1 mentions)
Huvec line, by PromoCell (1 mentions)
Gentamycin, by Merck Group (1 mentions)
Collagen type 1, by Advanced BioMatrix (1 mentions)
Amaxa huvec nucleofector kit, by Lonza (1 mentions)
Amphotericin b, by Harvard Bioscience (1 mentions)
Medium 199, by Merck Group (1 mentions)
Human umbilical vein endothelial cells (huvec), by PromoCell (1 mentions)
2 protocols using endothelial cell growth medium 2 supplementmix
1
Culturing Human Umbilical Vein Endothelial Cells
2
Culturing and Transfecting HUVECs
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Human umbilical vein endothelial cells (HUVEC) were obtained from Promocell and maintained as previously described (Heitzig et al, 2017) . were cultured in HUVEC medium composed of 50% Endothelial Cell Growth Medium 2 with Endothelial Cell Growth Medium 2 SupplementMix (C-39216, PromoCell) and 50% Medium 199 (Sigma) supplemented with 10% Fetal Bovine Serum Advanced (Capricorn Scientific), 30 µg/mL gentamycin (Sigma) and 15 ng/mL amphotericin B (Biochrom) at 5% CO2 and 37 °C on Corning CellBIND culture dishes. Coverslips and multiwell plates were coated with collagen type I (Advanced Biomatrix). Cells were transfected using the Amaxa HUVEC Nucleofector kit (Lonza) according to the protocol (2-5 µg plasmid DNA, 400 pmol siRNA duplex per 3x10 6 cells) and were analyzed 48 h post transfection.
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